Preparation method and application of Ni-NTA modified silicon dioxide coated ferroferric oxide magnetic nano functional assembly

A technology of ferroferric oxide and magnetic nanometers, which is applied in the field of materials, can solve the problems of complex preparation process, low separation efficiency, and insufficient surface group density, and achieve the effect of simple process, improved overall performance, and a large number of separation and extraction

Active Publication Date: 2020-05-19
甘肃省科学院传感技术研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a method for preparing a magnetic nano-functional assembly to sol

Method used

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  • Preparation method and application of Ni-NTA modified silicon dioxide coated ferroferric oxide magnetic nano functional assembly
  • Preparation method and application of Ni-NTA modified silicon dioxide coated ferroferric oxide magnetic nano functional assembly
  • Preparation method and application of Ni-NTA modified silicon dioxide coated ferroferric oxide magnetic nano functional assembly

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] (1) Fe 3 o 4 Preparation of nanoclusters:

[0038] Weigh 0.34gFeCl 3 ·6H 2 O, 0.9g NaAc 3H 2 Dissolve O in 60mL of ethylene glycol, stir at room temperature to completely dissolve the solid, and obtain a mixed solution; add the mixed solution into a 100mL stainless steel high-temperature reaction kettle embedded with polytetrafluoroethylene, react at 180°C for 8 hours, and use it after cooling Washed three times with ethanol and deionized water, separated by an external magnetic field, and freeze-dried to obtain Fe 3 o 4 nanoclusters;

[0039] Figure 5 In: (a) Fe 3 o 4 ; (b) Fe 3 o 4 @SiO 2 ; (c) Ni-NTA modified Fe 3 o 4 @SiO 2 TEM photographs of linked proteins in magnetic nanofunctional assemblies;

[0040] From Figure 5 (a) Fe can be seen in the transmission electron microscope 3 o 4 Nanoclusters are composed of many small Fe 3 o 4 Nanoparticles are assembled, and the cluster particle size is about 200nm.

[0041] From figure 2 (1) Fe can be...

Embodiment 2

[0064] As described in Example 1, the difference is:

[0065] Step (1) Fe 3 o 4 The preparation of nanoclusters remains unchanged;

[0066] Step (2) Fe 3 o 4 @SiO 2 The preparation remains unchanged;

[0067] In step (3), 0.5 mL of sodium N-(trimethoxysilylpropyl)ethylenediaminetriacetate was added, the heating temperature was 60° C., and the heating time was 10 h;

[0068] Add 10mgEDC and 10mgNHS in step (4), add 1mg NTA, the reaction time is 48h;

[0069] Add 0.1MNiCl in step (5) 2 Aqueous solution, mixed reaction 3h.

Embodiment 3

[0071] As described in Example 1, the difference is:

[0072] Step (1) Fe 3 o 4 The preparation of nanoclusters remains unchanged;

[0073] Step (2) Fe 3 o 4 @SiO 2 The preparation remains unchanged;

[0074] In step (3), 2 mL of sodium N-(trimethoxysilylpropyl) ethylenediamine triacetate was added, the heating temperature was 70° C., and the heating time was 9 h;

[0075] Add 20mgEDC and 20mgNHS in step (4), add 2mg NA, NA-bis(carboxymethyl)-L-lysine hydrate, and the reaction time is 36h;

[0076] Add 0.5MNiCl in step (5) 2 Aqueous solution, mixed reaction 2.5h.

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Abstract

The invention discloses a preparation method and applications of a Ni-NTA modified silicon dioxide coated ferroferric oxide magnetic nano functional assembly, and belongs to the field of materials. The preparation method comprises the following steps: preparing Fe3O4 nanoclusters by a solvothermal method; preparing Fe3O4@SiO2 magnetic nanoparticles by a St-ber sol-gel method; adding a silane coupling agent, and carrying out surface carboxylation of Fe3O4@SiO2 (Fe3O4@SiO2@COOH) through a chemical method; introducing an activating agent, and completing NTA modification (Fe3O4@SiO2@COOH@NTA) on the surfaces of the carboxylated Fe3O4@SiO2 magnetic nanoparticles; and adding Ni<2+> to prepare the assembly. According to the invention, the steps are clear, and the prepared magnetic nano functionalassembly is rich in surface groups, large in Ni<2+> binding capacity, high in binding force with His tag protein and high in separation efficiency.

Description

technical field [0001] The invention belongs to the field of materials, and in particular relates to a method for preparing a magnetic functional assembly for separating and extracting His-tagged proteins. Background technique [0002] With the development of biological science and biotechnology, methods for separating proteins and peptides have become diverse, including precipitation, ultrafiltration, chromatography (size-exclusion chromatography, ion-exchange chromatography, affinity chromatography, etc.), two-dimensional electrophoresis Wait. A good separation method should have the following three advantages: first, the coupling technology has high selectivity, good stability, and reproducibility; second, the protein load is high, controllable, reproducible, and easy to detect; third, It has the ability to resist non-specific protein adsorption and keep protein activity unaffected. [0003] With the research and development of magnetic nanoparticles, based on its stron...

Claims

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Application Information

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IPC IPC(8): B01J20/22B01J20/28B01J20/32C07K1/22B01D15/08
CPCB01D15/08B01J20/06B01J20/103B01J20/223B01J20/28009B01J20/3246C07K1/22
Inventor 冯治棋左显维刘一丹韩根亮李工农王玉祥何永成何欣
Owner 甘肃省科学院传感技术研究所
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